Antibiotic Resistance Protocols (Methods in Molecular Biology)

43

2. Cryostat to produce 3–5 μm thick frozen tissue sections (see
   Note 1).


3. Formalin solution.


4. Sucrose.


5. PBS.


6. Quartz coverslip (01015T-AB, SPI Supplies, PA, USA).


7. Quartz slide (01016-AB, SPI Supplies, PA, USA).


8. Transparent nail polish.

An example of the Raman system design is presented in Fig. 1.

1. Tunable laser that can generate light at 785 nm (see Note 2).


2. Microscope objective (Nikon, 50×, oil immersion).


3. Spectrometer: formed with a monochromator (Andor
   Shamrock SR303i, 400 lines/mm grating at 850 nm) and a
   cooled CCD camera (Newton 920, Andor Technology, UK).


4. A computer linked to the Ramen system and connected to the
   Internet.

2.3 Raman System

Fig. 1 Schematic diagram of the experimental setup. The system is using a tun-

able Ti:Sapphire laser (Spectra-Physics 3900 s, 785 nm, 1 W) pumped by a green

laser (Verdi V6, 532 nm, 5 W). L 1 , L 2 , and L 3 are lens. LF denotes line filter. EF

denotes edge filter and NF denotes notch filter. The laser is focused on the sam-

ple using a microscope objective (Nikon, 50×, oil). The spectrometer is com-

posed of a monochromator (Andor Shamrock SR303i, 400 lines/mm grating at

850 nm) and a cooled CCD camera (Andor Newton)

Detecting Phenotypically Resistant M. tb